N3,N6-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamide and use thereof in the field of neurotoxicity

ABSTRACT

The invention relates to a molecule enabling removal of neurotoxicity observed in neuron cells due to various reasons.

THE RELATED ART

The invention relates to a molecule enabling removal of neurotoxicityobserved in neuron cells due to various reasons.

The invention particularly relates to theN³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamidecompound obtained by using two different Nifepidine and Melatoninmolecules as the starting materials.

THE PRIOR ART

Brain is an organ that is almost impossible to recover functionallyfollowing serious neuronal damages. Damaged neurons either die or healslowly in a certain time depending on the developmental age of theorganism, anatomic localization of the cell, and the type, severity, andtime period of the damage.

Normally, the glutamate concentration released to the synaptic gap canreach very high levels; but such a high concentration only lasts for afew milliseconds. When this period is extended, neurons are exposed to akiller excitation due to excessive stimulation of neuronal glutamatereceptors. As a result of studies researching the effects of hypoxia andcerebral impacts, it was found out that exitotoxicity is responsible forthe destruction encountered after cerebral damage. In experimentalanimal studies, ischemia-related neuronal damage can be prevented viamicroinjection of glutamate receptor antagonists. It is reported thatglutamate antagonists are neuroprotective in stroke and head trauma inrodents and primates. All these results show that; intensiveaccumulation of glutamate during ischemia stimulates glutamatereceptors, which triggers a series of reactions causing neuronal death.It is believed that the reduced oxygen concentration as a result ofischemia prevents energy dependent glutamate re-uptake and thus causeincrease in glutamate.

Glaucoma is reported as the worldwide most common reason forirreversible vision loss. It is believed to effect more than fiftymillion people. While the reason for retina ganglion cell (RGH) losscannot be understood completely; the major risk factor causing glaucomais high intraocular pressure. The prior art treatments for glaucomaensure reduction of intraocular pressure via medicine or surgicaltechniques. However, pressure-independent loss of field of visioncontinues in some patients even though the intraocular pressure isreduced. It is known that in glaucomatous optic neuropathy also theprocess of formation of pathology is multifactorial like in otherneurodegenerative diseases, and variable cellular factors play a role inoptic neuropathy in molecular level. Any treatment mechanism thatdelays, protects, or reverses neuron cell death is called as “neuronprotection”.

Two important situations that cause neuronal damage in case of energydeficiency are disruption of the reabsorption of excitatory amino acidglutamate that is released to the synaptic gap, and the function loss ofthe sodium-potassium pump required for increase of glutamate in thesynaptic gap and maintenance of the membrane potential. Loss of cellmembrane potential as well as stimulation of NMDA and AMPA receptors bythe glutamate accumulated in the synaptic gap increase sodium andcalcium permeability of the cell. As a result intracellular sodium andcalcium concentration increase. It is known that permanent brain damageis reduced if the event is stopped at this stage. Most neuroprotectiveagents used in experimental studies are effective in techniques up tothis stage.

The effects of the prior art treatments on glutamate toxicity arelimited. Early treatment is essential for these diseases and the priorart treatments extend the treatment period. This negatively affects theprogression of disease.

As a result, the above said drawbacks and the inadequacy of the priorart solutions about the subject have necessitated an improvement in thetechnical field related to compounds eliminating neurotoxicity observedin neuron cells due to various reasons.

PURPOSE OF THE INVENTION

The invention relates to a molecule for eliminating neurotoxicityobserved in neuron cells due to various reasons, which meets the abovesaid requirements, eliminates all of the drawbacks, and brings someadditional advantages.

The primary purpose of the invention is to be a molecule whicheliminates neurotoxicity observed in neuron cells due to variousreasons.

A purpose of the invention is to be a specific glutamate blocker.

Another purpose of the invention is to have anti-oxidantcharacteristics.

Another purpose of the invention is to be a T-type Ca channel blocker.

In order to achieve the above said purposes, the invention comprisesN³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamidemolecule with the below given formulation:

which eliminates neurotoxicity observed in neuron cells.

The structural and characteristic features of the invention and all ofits advantages shall be understood better with the figures and thedetailed description given below in reference to the figures, andtherefore, the assessment should be made by taking into account the saidfigures and detailed explanations.

DETAILED DESCRIPTION OF THE INVENTION

In this detailed description, the preferred embodiments of a moleculefor eliminating neurotoxicity observed in neuron cells due to variousreasons, according to the invention are only disclosed for betterunderstanding of the subject without forming any limiting effect.

The invention is a molecule which eliminates neurotoxicity observed inneuron cells due to various reasons. The chemical compound according tothe invention is:N³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamide.

The molecule according to the invention defines two different functions,since it is formed of combination of two different molecules. Saidmolecules are: Nifepidine and Melatonin.

Nifepidine is a substance having the structure of calcium-channelblocker dihydropyridine. It selectively inhibits entrance of calciumions into the cell even at low doses, by being attached to calciumchannels.

Melatonin spreads extensively along the organism and enters cellularcompartments easily due to its amphophilic and small molecularstructure. This natural compound is a strong in vitro and in vivocytostatic agent with strong antioxidant characteristics.

The Synthesis Method ofN³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamideMolecule According to the Invention

Synthesis of 5-methoxytryptamine (2): Melatonin (1) is Used as theStarting Material of Said Synthesis

Melatonin is boiled at 90° C. for 8 hours in a 1 g (4.3 mmol) 40 mL 10%H₂SO₄ (sulphuric acid) solution. After the reaction is brought to roomtemperature, 20% of NaOH (sodium hydroxide) is added until the mediumbecomes alkaline. Afterwards, it is washed with ethyl acetate (3×20) andthe organic phase is dried over Na₂SO₄. The solvent is removed undervacuum. 550 mg crude product is obtained (67%) as a yellow solid.

Synthesis of dimethyl2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylate (4): Nifepidine(3) is Used as the Starting Material of Said Synthesis After nifepidineis dissolved in 1 g (2.89 mmol) 25 mL CH₂Cl₂(Dichloromethane), 1.37 g(3.18 mmol) PIFA ((Bis(trifluoroacetoxy)iodo)benzene) is added. Themixture is stirred at room temperature for 12 hours. Crude product iswashed with water (3×20) and the organic phase is dried over Na₂SO₄. Thesolvent of the organic phase is removed under vacuum. Oxidation productis crystallized over hexane. (900 mg, 90%). Synthesis of2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylic acid (5):Dimethyl 2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylate (4)Obtained in the Previous Step is Used as the Starting Material of SaidSynthesis

After dissolving dimethyl2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylate in 900 mg (2.61mmol) 15 mL ethanol and 15 mL H₂O, 733 mg (13.07 mmol) KOH is added. Thereaction is boiled at 80° C. for 16 hours. After the reaction is broughtto room temperature, its solvent is removed under vacuum. Afterwards, itis dissolved in 30 mL water, and 0.1 M HCl is added to adjust the pH to2, and it is kept at 2° C. for 12 hours. Obtained crystals are left todrying after filtering. 50 mg (66%) pure product is obtained.

Synthesis ofN³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamide(6): Dimethyl 2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylate(4) Obtained in the Previous Step and 5-methoxytryptamine (2) Obtainedin the First Step are Used as the Starting Materials of Said Synthesis

After dissolving 2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxylicacid in 550 mg (1.74 mmol) 10 mL THF (Tetrahydrofuran), HOBt(1-Hydroxybenzotriazole hydrate) 470 mg (3.488 mmol) is added and mixedfor 10 minutes. Subsequently, 661 mg (3.48 mmol) DCC is added to thereaction mixture and stirred for 30 minutes (1st mixture).

In another reaction balloon, after dissolving 5-methoxytryptamine (2) in661 mg (3.48 mmol) 10 mL THF, Et₃N (Triethylamine) 485 μL (3.48 mmol) isadded and stirred for 30 minutes (2nd mixture).

The resulting 2nd mixture is added to the 1st mixture, and stirred atroom temperature for 12 hours. Afterwards, the DCU formed is filteredthrough filter paper, and the solvent is removed under vacuum. Crudeproduct is firstly washed with NaHCO₃ (3×30 mL); and then with 5% KHSO₄(3×30 mL) and finally with water (3×30 mL), and the organic phase isdried over Na₂SO₄. The solvent of the organic phase is removed undervacuum. The resulting crude product is purified through silica gelcolumn with MeOH/CH₂Cl₂ (5%) and 400 mg (35%) yellow solid product isobtained.

The active regions of the compound of the invention are marked as shownbelow. A characteristic of the invention is blocking the Ca⁺⁺ channelstogether with the dihydropyridine group and by showing antioxidanteffect with the help of the carbonyl group, providing protective effectagainst the glutamate toxicity of the cell.

The Raw Materials Used During Synthesis of the Molecule of the Inventionwith the Above Given Operation Steps and their % Ratios:

Raw material Preferred amount by weight (%) Nifepidine 20 Melatonin 20H₂SO₄ 1.331 NaOH 2 Ethyl acetate 4 Na₂SO₄ 4 PIFA 1.37 H₂O 15 Hexane 2Ethanol 6 KOH 0.733 HCl 0.1M 6 THF 4 HOBt 0.470 DCC 0.661 Et₃N 0.485NaHCO₃ 4 5% KHSO₄ 8

The medicine according to the invention can be administered orally or inthe form of an IV sustained release implant.

The invention claimed is:
 1. A compound of N³,N⁵-bis(2-(5-methoxy-1H-indole-3-yl)ethyl)-2,6-dimethyl-4-(2-nitrophenyl)pyridine-3,5-dicarboxamideof the following formula:

which eliminates neurotoxicity observed in neuron cells.